Clonal expansion of genome-intact HIV-1 in functionally polarized Th1 CD4+ T cells
Clonal expansion of genome-intact HIV-1 in functionally polarized Th1 CD4+ T cells
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DOI：
10.1172/jci93289
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发表时间：
2017-06-30
影响因子：
15.9
通讯作者：
Lichterfeld, Mathias
中科院分区：
文献类型：
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作者：
Lee, Guinevere Q.;Orlova-Fink, Nina;Lichterfeld, Mathias
由于HIV-1在CD4(+) T细胞中持续存在，导致慢性无法治愈的疾病，CD4(+) T细胞含有复制能力强的前病毒，但表现出很少或没有活性病毒基因表达，并有效抵抗抗逆转录病毒联合治疗（cART）。这些潜伏感染的T细胞在所有循环CD4(+) T细胞中只占极小的比例，但具有显著的长期稳定性，并且通常在整个生命中持续存在，原因尚不完全清楚。在这里，我们对来自未分离的外周血单个核细胞、离体分离的CD4(+) T细胞和功能极化记忆CD4(+) T细胞亚群的HIV-1 DNA进行了大量的单基因组、近全长的下一代测序。这种方法从cart治疗的个体中鉴定出多组独立的、接近全长的前病毒序列，这些序列完全相同，与携带完整HIV-1的CD4(+) T细胞的克隆扩增一致。来自这种克隆扩增的CD4(+) T细胞的完整的、接近全基因组的HIV-1 DNA序列构成了记忆CD4 T细胞中所有分析的基因组完整序列的62%，优先在th1极化细胞中观察到，在长达5年的时间内被纵向检测到，并且完全具有复制和感染能力。总之，这些数据表明，编码完整HIV-1的th1极化CD4(+) T细胞的克隆增殖代表了稳定潜伏感染CD4(+) T细胞库的驱动力。
HIV-1 causes a chronic, incurable disease due to its persistence in CD4(+) T cells that contain replication-competent provirus, but exhibit little or no active viral gene expression and effectively resist combination antiretroviral therapy (cART). These latently infected T cells represent an extremely small proportion of all circulating CD4(+) T cells but possess a remarkable long-term stability and typically persist throughout life, for reasons that are not fully understood. Here we performed massive single-genome, near-full-length next-generation sequencing of HIV-1 DNA derived from unfractionated peripheral blood mononuclear cells, ex vivo-isolated CD4(+) T cells, and subsets of functionally polarized memory CD4(+) T cells. This approach identified multiple sets of independent, near-full-length proviral sequences from cART-treated individuals that were completely identical, consistent with clonal expansion of CD4(+) T cells harboring intact HIV-1. Intact, near-full-genome HIV-1 DNA sequences that were derived from such clonally expanded CD4(+) T cells constituted 62% of all analyzed genome-intact sequences in memory CD4 T cells, were preferentially observed in Th1-polarized cells, were longitudinally detected over a duration of up to 5 years, and were fully replication-and infection-competent. Together, these data suggest that clonal proliferation of Th1-polarized CD4(+) T cells encoding for intact HIV-1 represents a driving force for stabilizing the pool of latently infected CD4(+) T cells.